Laser Biopsy Inc, (LBI) has a vision of improving the speed and quality of cancer diagnosis with reduced patient risk and reduced cost. LBI is developing a new medical imaging system that will enable pathologists to perform real-time diagnosis of biopsies taken during surgical procedures. Most cancer diagnoses are made by biopsying the suspicious lesions under some form of image guidance, followed by histopathological analysis of the fixed and stained tissue. However, with more effective screening strategies, disease is being diagnosed earlier, and in such cases, cancer is often not clearly distinguishable by imaging. Under these conditions, the surgeon operates relatively "blind" during the biopsy procedure, removing a certain number of random biopsies from "representative sites" within the organ, without the benefit of any real-time feedback to target the areas of the organ with cancer. In order to address this major gap in current state-of-the-art cancer care, Laser Biopsy Inc. is developing a compact low-cost Multiphoton microscopy (MPM) system to enable rapid nondestructive analysis of tissue samples within a minute of obtaining a biopsy specimen, without the need for any fixation chemicals, dye stains, or microtome sectioning. Multiphoton-based imaging technology allows visualization of tissue, providing information both about tissue architecture and cellular morphology. It is our overarching hypothesis that multiphoton histopathology from tissue biopsies can provide real-time feedback to surgeons, thus improving the accuracy of targeted biopsies and improving patient diagnosis and treatment. This project is aimed toward the development and demonstration of a novel, compact optical design for a low cost multiphoton microscope with the performance parameters of a high end commercial system. The LBI design has the ability to image in a single scan an area 25 times that of a conventional multiphoton microscope system without sacrificing resolution or the need for "tiling" many images together. Furthermore, the design makes use of the latest optical rapid prototyping technology and requires a minimal number of components for reduced cost and complexity. Based on current parts estimates, a selling price of sub $100K for the entire instrument is supportable, which amounts to a 3-6 x reduction in cost over current commercial systems. The system will be evaluated and tested against a commercial multiphoton microscope using both standardized fluorescent samples and animal tissue. The performance will be quantified by directly comparing resolution, signal intensity, penetration depth, and imaging time per equivalent section. The data will be reviewed by a surgeon and pathologist from the Veterinary School at Cornell. The work will form the basis for a phase II program, in which the results will be used to develop an "instant" pathology instrument for freshly excised tissue in time-critical, surgical procedures. PUBLIC HEALTH RELEVANCE: A compact, cost-effective, high performance multiphoton microscope will enable the visualization of freshly excised tissue with sub micron resolution, without fixation, sectioning or staining. Such an imaging modality will be used to provide immediate feedback to surgeons performing biopsies with improved quality of diagnosis, reduced cost per section, reduced operating room costs, reduced patient anxiety, and intrinsic archival digital records.